Generation of Monoclonal Antibodies Against Mouse Neurogenin 3: A New Immunocytochemical Tool to Study the Pancreatic Endocrine Progenitor Cell

Zahn, Stefan; Hecksher-Sørensen, Jakob; Pedersen, Inger L.; Serup, Palle; Madsen, Ole

doi:10.1089/1536859042729928

Cited by 6 publications

(6 citation statements)

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“…Primary antibodies: rabbit anti--galactosidase (MP Biomedicals), 1:1600; mouse anti-Neurog3 (Zahn et al, 2004), 1:500; goat anti-Pdx1 (Beta Cell Biology Consortium), 1:15,000; goat anti-GFP (Abcam), 1:1000; rabbit anti-Ptf1a (Hald et al, 2008), 1:4000.…”

Section: Immunohistochemistrymentioning

confidence: 99%

Ptf1a-mediated control of Dll1 reveals an alternative to the lateral inhibition mechanism

et al. 2012

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There was an error published in Development 139, 33-45.On p. 36, Fig. 1 was incorrectly cited several times in place of Fig. 2. The correct paragraph appears below.The authors apologise to readers for this mistake.To determine if endodermal Notch activation and Hes1 expression depends on Dll1 activity, we analyzed NICD expression in wild-type and Dll1 lacZ/lacZ embryos and EGFP expression in crosses of Tg(Hes1-EGFP) 1Hri and Dll1 lacZ/+ mice. NICD expression was reduced in E10.5 Dll1 lacZ/lacZ embryos compared with controls ( Fig. 2A,B), but appeared to recover, approaching wild-type levels at E11.5 (Fig. 2C,D). Hes1-EGFP expression was normal in E8.25 Dll1 lacZ/lacZ embryos (Fig. 2E,I) but had partly disappeared from the dorsal pancreas endoderm at E9.5 (Fig. 2F,J) and was almost lost at E10.5 (Fig. 2G,K). Remarkably, and coinciding with the reappearance of NICD, Hes1-EGFP expression was restored in E11.5 Dll1 lacZ/lacZ embryos (Fig. 2H,L).

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Section: Immunohistochemistrymentioning

confidence: 99%

Ptf1a-mediated control of Dll1 reveals an alternative to the lateral inhibition mechanism

et al. 2012

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“…Because of the light fixation, endocrine hormone signals appeared diffuse, and we could not identify the exact islet cell type(s) that express Neurog3 by using the guinea pig antibody. We also examined Neurog3 production in the adult islet cells by using a mouse monoclonal Neurog3 antibody (22) and a rabbit anti-Neurog3 (2). Both antibodies produced high background in lightly fixed postnatal islets, making it impossible to discern whether a true Neurog3 signal exists.…”

Section: Neurog3 Mrna and Protein Can Be Detected In Hormone-expressingmentioning

confidence: 99%

Sustained Neurog3 expression in hormone-expressing islet cells is required for endocrine maturation and function

Wang¹,

Jensen

Seymour

et al. 2009

Proc. Natl. Acad. Sci. U.S.A.

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Neurog3 (Neurogenin 3 or Ngn3) is both necessary and sufficient to induce endocrine islet cell differentiation from embryonic pancreatic progenitors. Since robust Neurog3 expression has not been detected in hormone-expressing cells, Neurog3 is used as an endocrine progenitor marker and regarded as dispensable for the function of differentiated islet cells. Here we used 3 independent lines of Neurog3 knock-in reporter mice and mRNA/protein-based assays to examine Neurog3 expression in hormone-expressing islet cells. Neurog3 mRNA and protein are detected in hormoneproducing cells at both embryonic and adult stages. Significantly, inactivating Neurog3 in insulin-expressing ␤ cells at embryonic stages or in Pdx1-expressing islet cells in adults impairs endocrine function, a phenotype that is accompanied by reduced expression of several Neurog3 target genes that are essential for islet cell differentiation, maturation, and function. These findings demonstrate that Neurog3 is required not only for initiating endocrine cell differentiation, but also for promoting islet cell maturation and maintaining islet function.endocrine progenitor ͉ maintenance ͉ pancreas ͉ diabetes ͉ sugar metabolism

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“…1B,C). Next we performed immunohistochemical analysis with the well-characterized F25A1B3 antibody (Zahn et al, 2004) to investigate the distribution of the protein in the mature mouse brain. Ngn3 was detected in all representative brain regions examined and, in agreement with the results obtained with in situ hybridization, immunohistochemical analysis showed that in the hippocampal formation Ngn3 is expressed in the stratum pyramidalis of fields CA1-3 and in the granular layer of the DG and in cells located in the neuropil of CA1 and the molecular layer of the dentate gyrus (Fig.…”

Section: Ngn3 Mrna and Protein Expression Pattern In Adult Hippocampusmentioning

confidence: 99%

Neurogenin 3 cellular and subcellular localization in the developing and adult hippocampus

Simon-Areces

Membrive

García-Fernández

et al. 2010

J of Comparative Neurology

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Neurogenin 3 (Ngn3), a proneural gene controlled by the Notch receptor, is implicated in the control of dendrite morphology and synaptic plasticity of cultured hippocampal neurons. Here we report the localization and subcellular distribution of Ngn3 in the hippocampus in vivo and in neuronal cultures. In situ hybridization showed Ngn3 mRNA expression in the pyramidal layer and dentate gyrus of adult mouse hippocampus. Immunohistochemistry studies revealed that Ngn3 localization is mostly cytoplasmic in the hippocampal eminence at embryonic day (E)17 and postnatal day (P)0. At P10 it is cytoplasmic in CA1-CA3 pyramidal neurons and nuclear in granule cells of the dentate gyrus. In the adult hippocampus Ngn3 is localized in the nucleus and cytoplasm of both pyramidal neurons and granule cells. During development of cultured hippocampal neurons, Ngn3 mRNA expression is higher at stages of neuronal polarization, as judged by reverse-transcription polymerase chain reaction (RT-PCR), and it is mostly cytoplasmic. The tracking of the subcellular localization of Ngn3 in neurons infected with a virus expressing myc-Ngn3 suggests that the protein is quickly translocated to the cell nucleus after synthesis and then reexported to the cytoplasm. Treatment with leptomycin B, a potent and specific inhibitor of the exportin CRM1, induced its accumulation into the nucleus, suggesting that CRM1 mediates the nuclear export of Ngn3. These results suggest that Ngn3 may play a role in neuronal development by actions in the cytoplasm.

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Generation of Monoclonal Antibodies Against Mouse Neurogenin 3: A New Immunocytochemical Tool to Study the Pancreatic Endocrine Progenitor Cell

Cited by 6 publications

References 0 publications

Ptf1a-mediated control of Dll1 reveals an alternative to the lateral inhibition mechanism

Ptf1a-mediated control of Dll1 reveals an alternative to the lateral inhibition mechanism

Sustained Neurog3 expression in hormone-expressing islet cells is required for endocrine maturation and function

Neurogenin 3 cellular and subcellular localization in the developing and adult hippocampus

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